SERGAS, Research Laboratory 9 (NEIRID LAB: Neuroendocrine Interactions in Rheumatology and Inflammatory Diseases), Institute of Medical Research (IDIS), Santiago University Clinical Hospital, Santiago de Compostela, 15706, Spain.
J Orthop Res. 2014 May;32(5):653-60. doi: 10.1002/jor.22585. Epub 2014 Jan 25.
Nesfatin-1 is a recently discovered satiety-inducing adipokine identified in hypothalamic regions that regulates energy balance. So far, no data exist on NUCB2/nesfatin-1 localization in human and murine chondrocytes. Here, we therefore investigated NUCB2/nesfatin-1 gene and protein expression in human and murine chondrocytes and the effect of nesfatin-1 on pro-inflammatory cytokines expression. Peptide localization was performed by laser confocal microscopy, NUCB2 mRNA expression was studied by RT-PCR and protein secretion was measured by XMap technology and Western blot analysis. First, we demonstrated cytoplasmic localization of NUCB2/nesfatin-1 peptide in both human and murine chondrocytes. We present evidence that both mRNA and protein expression of NUCB2 were increased during the differentiation of ATDC5 murine chondrocyte cell line. Furthermore, we demonstrated that nesfatin-1 induces IL-6 and MIP-1α mRNA expression and protein secretion in ATDC-5 cells challenged with IL-1, and also increases COX-2 mRNA expression in these cells. Finally, nesfatin-1 provoked a clear induction of pro-inflammatory agents, such as COX-2, IL-8, IL-6, and MIP-1α in human primary chondrocytes from OA patients.
内脂素-1 是一种最近发现的具有饱腹感的脂肪因子，存在于下丘脑区域，调节能量平衡。到目前为止，还没有关于 NUCB2/内脂素-1 在人及鼠类软骨细胞中的定位的数据。因此，本研究旨在检测人及鼠类软骨细胞中 NUCB2/内脂素-1 基因和蛋白的表达，并研究内脂素-1 对促炎细胞因子表达的影响。通过激光共聚焦显微镜检测肽的定位，通过 RT-PCR 检测 NUCB2mRNA 的表达，通过 XMap 技术和 Western blot 分析检测蛋白分泌。首先，我们证实了内脂素-1 肽在人及鼠类软骨细胞的细胞质中存在定位。本研究证明，在 ATDC5 鼠类软骨细胞系分化过程中，NUCB2 的 mRNA 和蛋白表达均增加。此外，我们发现内脂素-1 可诱导 ATDC-5 细胞中 IL-1 诱导的 IL-6 和 MIP-1αmRNA 表达和蛋白分泌增加，并且还可增加这些细胞中的 COX-2 mRNA 表达。最后，内脂素-1 可明显诱导 OA 患者的人原代软骨细胞中促炎介质（如 COX-2、IL-8、IL-6 和 MIP-1α）的产生。
